# 95: DESULFUROMONAS ACETOXIDANS MEDIUM


## Main sol. 95

951	ml	**Solution A**	
3	ml	**Solution B**	
40	ml	**Solution C**	
10	ml	**Solution D**	

1. Sparge solution A with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense solution under same gas atmosphere into anoxic Hungate-type tubes or serum vials and sterilize by autoclaving. Solutions B and D are autoclaved separately under 100% N2 gas atmosphere. Solution C is autoclaved under 80% N2 and 20% CO2 gas atmosphere. The medium is completed before use by adding appropriate amounts of solutions B, C and D to solution A. Adjust pH of complete medium to 7.2, if necessary.


## Solution A

1	g	KH2PO4	
0.3	g	NH4Cl	
1	g	MgSO4 x 7 H2O	
2	g	MgCl2 x 6 H2O	
20	g	NaCl	
0.1	g	CaCl2 x 2 H2O	
1.77	g	Na2SO4	
1	ml	**Trace element solution SL-11**	
2.66	g	Na2-DL-malate	
0.5	g	Yeast extract	
0.5	ml	Sodium resazurin (0.1% w/v)	
950	ml	Distilled water	

1. Adjust pH to 6.0 with 2 N NaOH.


## Solution B

0.3	ml	Ethanol	
2.7	ml	Distilled water	


## Solution C

1.85	g	NaHCO3	
40	ml	Distilled water	


## Solution D

0.3	g	Na2S x 9 H2O	
10	ml	Distilled water	


## Trace element solution SL-11

5.2	g	Na2-EDTA x 2 H2O	
1.5	g	FeCl2 x 4 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 ml.